CN111603942A - Protein separation membrane based on interface assembly mode, preparation method and application thereof - Google Patents
Protein separation membrane based on interface assembly mode, preparation method and application thereof Download PDFInfo
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- CN111603942A CN111603942A CN202010532762.1A CN202010532762A CN111603942A CN 111603942 A CN111603942 A CN 111603942A CN 202010532762 A CN202010532762 A CN 202010532762A CN 111603942 A CN111603942 A CN 111603942A
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D69/00—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor
- B01D69/02—Semi-permeable membranes for separation processes or apparatus characterised by their form, structure or properties; Manufacturing processes specially adapted therefor characterised by their properties
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
- B01D67/0002—Organic membrane manufacture
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D67/00—Processes specially adapted for manufacturing semi-permeable membranes for separation processes or apparatus
- B01D67/0002—Organic membrane manufacture
- B01D67/0006—Organic membrane manufacture by chemical reactions
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D71/00—Semi-permeable membranes for separation processes or apparatus characterised by the material; Manufacturing processes specially adapted therefor
- B01D71/06—Organic material
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B21/00—Thiazine dyes
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B29/00—Monoazo dyes prepared by diazotising and coupling
- C09B29/06—Monoazo dyes prepared by diazotising and coupling from coupling components containing amino as the only directing group
- C09B29/08—Amino benzenes
- C09B29/0801—Amino benzenes containing acid groups, e.g. COOH, SO3H, PO3H2, OSO3H, OPO3H2; SO2NHSO2R or salts thereof, R being hydrocarbonyls
- C09B29/0803—Amino benzenes containing acid groups, e.g. COOH, SO3H, PO3H2, OSO3H, OPO3H2; SO2NHSO2R or salts thereof, R being hydrocarbonyls containing SO3H, OSO3H
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B57/00—Other synthetic dyes of known constitution
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0096—Purification; Precipitation; Filtration
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- Manufacturing & Machinery (AREA)
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Abstract
The invention discloses a protein separation membrane based on an interface assembly mode, a preparation method and application thereof, belonging to the technical field of protein separation membranes. The separation membrane has the advantages of low cost, environmental protection, simple preparation process and the like, and has good biocompatibility. According to the characteristics of interface assembly, the size of the separation membrane is determined by the size of the interface, so that large-area preparation is easier to realize. The separation membrane has the advantages of selective separation of molecules with different charges, complete interception of molecules with negative charges and higher diffusion rate of small molecules with positive charges, and is an ideal separation membrane.
Description
Technical Field
The invention belongs to the technical field of protein separation membranes, and particularly relates to a protein separation membrane based on an interface assembly mode, a preparation method and application thereof.
Background
In recent years, membrane separation technology has been rapidly developed and is a major development project in many countries of the world.
As a high and new technology, membrane separation is widely applied in various fields such as environmental protection, water treatment, chemical industry, metallurgy, energy, food and the like. In the field of research on membrane separation, research on membrane materials is an important issue. At present, commonly used membrane materials are mainly organic synthetic polymer materials, however, the preparation method of the polymer membrane is complex, and inevitable limitations are brought in the preparation process, so that the separation performance of the biological membrane is influenced. On the other hand, because of its good inherent biocompatibility and biodegradability, biofilms have been a source of inspiration for the development of modern membrane materials.
Therefore, it is of great practical significance to develop a bio-separation membrane which has good separation performance, is simple and easy to implement, and is low in cost.
Disclosure of Invention
Aiming at the defects of complex polymer membrane preparation process and the like in the prior art, the invention provides a protein separation membrane based on an interface assembly mode, a preparation method and application of the separation membrane.
The invention is realized by the following technical scheme:
a protein separation membrane based on an interface assembly mode is a two-dimensional nano-film formed by self-assembly of Bovine Serum Albumin (BSA) and a surfactant after electrostatic interaction occurs at an oil-water interface.
Preferably, the surfactant is cetyltrimethylammonium bromide (CTAB).
Preferably, the thickness of the two-dimensional nano-film ranges from 3.2nm to 625 nm.
The invention also provides a preparation method of the protein separation membrane based on the interface assembly mode, which comprises the following specific steps:
the method comprises the following steps: preparing a PBS buffer solution containing Bovine Serum Albumin (BSA);
step two: mixing the PBS buffer solution containing Bovine Serum Albumin (BSA) prepared in the step one with a surfactant, and then adding a 2-ethyl-1-hexanol (isooctanol) solution, wherein the isooctanol forms an oil-water interface due to the insolubility with water;
step three: and (3) adding a glutaraldehyde solution into the mixed solution in the second step for crosslinking, and incubating at room temperature for 1-2 hours to observe a layer of BSA protein two-dimensional nano-film on the oil-water interface.
Preferably, the concentration of the PBS buffer solution containing Bovine Serum Albumin (BSA) in step one is 1-20 mg/ml.
Preferably, the PBS buffer solution containing Bovine Serum Albumin (BSA) in step one has a pH of 7.
Preferably, the volume ratio of the PBS buffer solution containing Bovine Serum Albumin (BSA) to the surfactant in step two is 1: 1
Preferably, the volume ratio of the 2-ethyl-1-hexanol to the mixed solution in the second step is 1: 4.
preferably, the surfactant in step two is cetyl trimethyl ammonium bromide.
Preferably, the mass fraction of the glutaraldehyde solution in step three is 2%.
The invention also provides an application of the protein separation membrane based on the interface assembly mode in separation of mixed dyes, wherein the mixed dyes are methylene blue and methyl orange, or methyl blue and rhodamine B.
Compared with the prior art, the invention has the following advantages:
1. the separating membrane prepared by the invention is formed by tightly packing BSA protein, is colorless and transparent, is a pure protein membrane, and can control the thickness of the membrane according to the concentration of the BSA protein.
2. The prepared separation membrane has good biocompatibility and biodegradability, and because the surface of the BSA protein separation membrane has strong positive charges, the BSA protein separation membrane can effectively intercept dyes with negative charges in a neutral environment, and the interception effect of dye molecules with positive charges is not obvious, so that the separation membrane can be effectively applied to selective separation of a mixed dye solution.
3. The separation membrane can effectively separate methylene blue, methyl orange, methyl blue and rhodamine B.
4. The preparation method of the separation membrane is simple, is easy to realize large-area preparation, has the characteristics of low cost, low energy consumption, environmental protection and the like, and avoids the problems of complicated steps and environmental pollution in the traditional polymer membrane synthesis process.
Drawings
In order to more clearly illustrate the detailed description of the invention or the technical solutions in the prior art, the drawings that are needed in the detailed description of the invention or the prior art will be briefly described below. Throughout the drawings, like elements or portions are generally identified by like reference numerals. In the drawings, elements or portions are not necessarily drawn to scale.
FIG. 1 is an atomic force microscope image of a cross-linked BSA protein film in example 2 of the present invention, wherein A is an atomic force plan view of the protein film, and B is an atomic force microscope height (i.e., film thickness) of the protein film;
FIG. 2 is a plan view of an optical microscope showing a cross-linked BSA protein film in example 1 of the present invention;
FIG. 3 is a plan view of a transmission electron microscope showing a cross-linked BSA protein film in example 1 of the present invention;
FIG. 4 is the separation of methyl blue and rhodamine B by the separation membrane obtained in example 1 of the present invention;
FIG. 5 shows the separation of methylene blue and methyl orange by the separation membrane obtained in example 1 of the present invention.
Detailed Description
The following embodiments are only used for illustrating the technical solutions of the present invention more clearly, and therefore, the following embodiments are only used as examples, and the protection scope of the present invention is not limited thereby.
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains.
The invention provides a protein separation membrane based on an interface assembly mode, which is a two-dimensional nano-film formed by self-assembling Bovine Serum Albumin (BSA) and a surfactant after electrostatic interaction occurs at an oil-water interface.
The surfactant is Cetyl Trimethyl Ammonium Bromide (CTAB).
The thickness range of the two-dimensional nano film is 3.2nm-625 nm.
Example 1
1ml of 2mg/ml BSA in PBS buffer (pH 7) and 1ml of 3mg/ml CTAB aqueous solution were mixed in equal volumes in a petri dish with a diameter of 3.5cm, then 400. mu.L of 2-ethyl-1-hexanol solution was added, then 25. mu.L of glutaraldehyde solution was added to the solution, and incubation was carried out at room temperature for 1-2 hours, and a layer of BSA protein two-dimensional nano-membrane was observed at the oil-water interface.
Example 2
In this example, a PBS buffer solution (pH 7) of 2mg/ml BSA and a 3mg/ml CTAB aqueous solution in example 1 were replaced with a PBS buffer solution (pH 7) of 4mg/ml lbs a and 1ml of a 6mg/ml CTAB aqueous solution at equal volumes, and the other steps were the same as in example 1 to obtain a separation membrane.
Example 3
In this example, a PBS buffer solution (pH 7) of 2mg/ml BSA and a 3mg/ml CTAB aqueous solution in example 1 were replaced with a PBS buffer solution (pH 7) of 6mg/ml lbs a and 1ml of 9mg/ml CTAB aqueous solution at equal volumes, and the other steps were the same as in example 1 to obtain a separation membrane.
Example 4
In this example, a PBS buffer solution (pH 7) of 2mg/ml BSA and a 3mg/ml CTAB aqueous solution in example 1 were replaced with a PBS buffer solution (pH 7) of 8mg/ml lbs a and 1ml of 12mg/ml CTAB aqueous solution at equal volumes, and the other steps were the same as in example 1 to obtain a separation membrane.
Example 5
In this example, a PBS buffer solution (pH 7) of 2mg/ml BSA and a 3mg/ml CTAB aqueous solution in example 1 were replaced with a PBS buffer solution (pH 7) of 10mg/ml lbs a and 1ml of 15mg/ml CTAB aqueous solution at equal volumes, and the other steps were the same as in example 1 to obtain a separation membrane.
Atomic force and scanning electron microscope characterization was performed on the crosslinked BSA protein nano-films prepared in examples 1-5, and the results showed that the corresponding film thicknesses were 3.2nm, 12nm (fig. 1), 32nm, 500nm, and 625nm in sequence, and the optical microscope image (fig. 2) and the scanning electron microscope image (fig. 3) showed the successfully assembled micron-sized protein films. The protein film has a high degree of flexibility and independence, as can also be seen from the folds and folded edges.
Example 6
The invention discloses an application of a protein separation membrane based on an interface assembly mode in mixed dye separation, wherein the mixed dye is methylene blue and methyl orange, or methyl blue and rhodamine B.
The specific method comprises the following steps:
the separation membrane prepared in example 1 was floated on 3mL of ultrapure water, 20. mu.L of an aqueous solution containing 50mg/L of methylene blue and 50mg/L of methyl orange and 25. mu.L of an aqueous solution containing 50mg/L of methyl blue and 50mg/L of rhodamine were dropped on the membrane, respectively, and after standing at room temperature for 12 hours, the solution was monitored for permeability by ultraviolet-visible absorption spectroscopy. Experiment results show that the separation membrane has strong positive charges, so that the dye with negative charges can be effectively intercepted in a neutral environment, the molecular interception effect of the dye with positive charges is not obvious, the interception rate of methyl blue is about 100%, the interception rate of rhodamine B is about 21%, the interception rate of methylene blue is about 1.4%, the interception rate of methyl orange is about 92%, and the quick and effective separation of the methyl blue and the rhodamine B (figure 4), the methylene blue and the methyl orange (figure 5) can be realized.
The preferred embodiments of the present invention have been described in detail with reference to the accompanying drawings, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
Claims (10)
1. The protein separation membrane based on the interface assembly mode is characterized in that the membrane is a two-dimensional nano-film formed by self-assembling Bovine Serum Albumin (BSA) and a surfactant after electrostatic interaction occurs at an oil-water interface.
2. The interfacial assembly based protein separation membrane according to claim 1, wherein the surfactant is cetyltrimethylammonium bromide.
3. The protein separation membrane according to claim 1, wherein the thickness of the two-dimensional nano-film is in the range of 3.2nm to 625 nm.
4. The method for preparing a protein separation membrane based on an interface assembly mode according to claim 1, which comprises the following steps:
the method comprises the following steps: preparing a PBS buffer solution containing Bovine Serum Albumin (BSA);
step two: mixing the PBS buffer solution containing Bovine Serum Albumin (BSA) prepared in the step one with a surfactant, and then adding a 2-ethyl-1-hexanol (isooctanol) solution, wherein the isooctanol forms an oil-water interface due to the insolubility with water;
step three: and (3) adding a glutaraldehyde solution into the mixed solution in the second step for crosslinking, and incubating at room temperature for 1-2 hours to observe a layer of BSA protein two-dimensional nano-film on the oil-water interface.
5. The method according to claim 4, wherein the concentration of the PBS buffer solution containing Bovine Serum Albumin (BSA) in the first step is 1-20 mg/ml; the PBS buffer solution containing Bovine Serum Albumin (BSA) has a pH value of 7.
6. The method according to claim 4, wherein the volume ratio of the PBS buffer solution containing Bovine Serum Albumin (BSA) to the surfactant in the second step is 1: 1; the volume ratio of the 2-ethyl-1-hexanol to the mixed solution is 1: 4.
7. the method according to claim 4, wherein the surfactant in step two is cetyltrimethylammonium bromide.
8. The method for preparing a protein separation membrane based on an interfacial assembly method according to claim 4, wherein the glutaraldehyde solution in step three is 2% by mass.
9. Use of the protein separation membrane based on the interfacial assembly method according to claim 1 in mixed dye separation.
10. The use of the interfacial assembly based protein separation membrane in the separation of mixed dyes according to claim 9, wherein the mixed dyes are methylene blue and methyl orange, or methyl blue and rhodamine B.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101862609A (en) * | 2007-07-11 | 2010-10-20 | 独立行政法人物质·材料研究机构 | Protein nano film, its manufacture method and the application of soft independence |
| CN103240006A (en) * | 2013-05-10 | 2013-08-14 | 天津大学 | Bovine serum albumin-polycarbonate composite membrane and preparation method thereof |
| CN108854599A (en) * | 2018-05-14 | 2018-11-23 | 陕西师范大学 | A kind of dialysis membrane and its application based on crosslinking lysozyme |
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2020
- 2020-06-12 CN CN202010532762.1A patent/CN111603942A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101862609A (en) * | 2007-07-11 | 2010-10-20 | 独立行政法人物质·材料研究机构 | Protein nano film, its manufacture method and the application of soft independence |
| CN103240006A (en) * | 2013-05-10 | 2013-08-14 | 天津大学 | Bovine serum albumin-polycarbonate composite membrane and preparation method thereof |
| CN108854599A (en) * | 2018-05-14 | 2018-11-23 | 陕西师范大学 | A kind of dialysis membrane and its application based on crosslinking lysozyme |
Non-Patent Citations (2)
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| FACUI YANG AT.AL: "Self-assembled membrane composed of amyloidlike proteins for eff", 《NATURE COMMUNICATIONS》 * |
| TINGTING WANG ET.AL: "Giant Proteinosomes As Scaffolds for Light Harvesting", 《AMERICAN CHEMICAL SOCIETY MACRO LETTERS》 * |
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Application publication date: 20200901 |